Secreted proteins of normal and myc-ras oncogene transformed rat embryo fibroblasts

Quiescent cultures of rat embryo fibroblasts synthesize and secrete several proteins in response to mitogenic stimulation. Two of these proteins have been characterized in this study and the effect of oncogenic transformation on these proteins was monitored. A serum induced 48,000 protein was shown to be related to plasminogen activator inhibitor while another serum-induced protein of Mr 45,000 was found to be an inhibitor of DNA synthesis. Transformation of rat embryo fibroblasts with oncogenesmyc andras resulted in drastic reduction in the level of these proteins. The reduced levels of protease inhibitor may be responsible for the loss of anchorage dependence of the transformed cells. The DNA synthesis inhibitor protein may act as a negative growth regulator and reduced levels of this protein in myc-ras transformed cells may accelerate the proliferation of these cells

A novel combinatorial technique for simultaneous quantification of oxygen radicals and aggregation reveals unexpected redox patterns in the activation of platelets by different physiopathological stimuli

This is the author accepted manuscript. The final version is available fromFerrata Storti Foundation via the DOI in this recordThe regulation of platelets by oxidants is critical for vascular health and may explain thrombotic complications in diseases such as diabetes and dementia, but remains poorly understood. Here, we describe a novel technique combining electron paramagnetic resonance spectroscopy and turbidimetry, which has been utilised to monitor simultaneously platelet activation and oxygen radical generation. This technique has been used to investigate the redox-dependence of human and mouse platelets. Using selective peptide inhibitors of NOXs on human platelets and genetically modified mouse platelets (NOX1-/- or NOX2-/-), we discovered that:1) intracellular but not extracellular superoxide anion generated by NADPH oxidases (NOXs) is critical for platelet activation by collagen; 2) superoxide dismutation to hydrogen peroxide is required for thrombin-dependent activation; 3) NOX1 is the main source of oxygen radicals in response to collagen, while NOX2 is critical for activation by thrombin; 4) two platelet modulators, namely oxidised low density lipoproteins (oxLDL) and amyloid peptide β (Aβ), require activation of both NOX1 and NOX2 to pre-activate platelets. This study provides new insights on the redox dependence of platelet activation. It suggests the possibility of selectively inhibiting platelet agonists by targeting either NOX1 (for collagen) or NOX2 (for thrombin). Selective inhibition of either NOX1 or NOX2 impairs the potentiatory effect of tested platelet modulators (oxLDL and Aβ), but does not completely abolish platelet haemostatic function. This information offers new opportunities for the development of disease specific antiplatelet drugs with limited bleeding side effects by selectively targeting one NOX isoenzyme.British Heart Foundatio

Common fixed point theorems of different compatible type mappings using Ciric\u27s contraction type condition

The purpose of this paper is to establish necessary and sufficient conditions for the existence of common fixed points for a compatible pair of selfmaps under Ciric\u27s contraction type condition. These theorems improve and generalize the results of Mukherjee and Verma [11] and Jungck [9] to a pair of selfmaps. Also established the existence of common fixed points for a pair of compatible mappings of type (B), and obtain a result on the existence of common fixed points for a pair of compatible mappings of type (A) as corollary. Greguš fixed point theorem follows as a special case to our results

A novel flow cytometry assay using dihydroethidium as redox-sensitive probe reveals NADPH oxidase-dependent generation of superoxide anion in human platelets exposed to amyloid peptide β

This is the author accepted manuscript. The final version is available from Taylor & Francis via the DOI in this recordReactive oxygen species (ROS) generation is critical in the regulation of platelets, which has important implications in the modulation of hemostasis and thrombosis. Nonetheless, despite several assays have been described and successfully utilized in the past, the analysis of ROS generation in human platelets remains challenging. Here we show that dihydroethidium (DHE) allows the characterization of redox responses upon platelet activation by physiological and pathological stimuli. In particular, the flow cytometry assay that we describe here allowed us to confirm that thrombin, collagen-related peptide (CRP) and arachidonic acid but not adenosine diphosphate (ADP) stimulate superoxide anion formation in a concentration-dependent manner. 0.1unit/ml thrombin, 3 μg/ml CRP and 30 μM arachidonic acid are commonly used to stimulate platelets in vitro and here were shown to stimulate a significant increase in superoxide anion formation. The ROS scavenger N-acetylcysteine (NAC) abolished superoxide anion generation in response to all tested stimuli, but the pan-NADPH oxidase (NOX) inhibitor VAS2870 only inhibited superoxide anion formation in response to thrombin and CRP. The involvement of NOXs in thrombin and CRP-dependent responses was confirmed by the inhibition of platelet aggregation induced by these stimuli by VAS2870, while platelet aggregation in response to arachidonic acid was insensitive to this inhibitor. In addition, the pathological platelet stimulus amyloid β (Aβ) 1–42 peptide induced superoxide anion formation in a concentration-dependent manner. Aβ peptide stimulated superoxide anion formation in a NOX-dependent manner, as proved by the use of VAS2870. Aβ 1–42 peptide displayed only moderate activity as an aggregation stimulus, but was able to significantly potentiate platelet aggregation in response to submaximal agonists concentrations, such as 0.03 unit/ml thrombin and 10 μM arachidonic acid. The inhibition of NOXs by 10 μM VAS2870 abolished Aβ-dependent potentiation of platelet aggregation in response to 10 μM arachidonic acid, suggesting that the pro-thrombotic activity of Aβ peptides depends on NOX activity. Similar experiments could not be performed with thrombin or collagen, as NOXs are required for the signaling induced by these stimuli. These findings shed some new light on the pro-thrombotic activity of Aβ peptides. In summary, here we describe a novel and reliable assay for the detection of superoxide anion in human platelets. This is particularly important for the investigation of the pathophysiological role of redox stress in platelets, a field of research of increasing importance, but hindered by the absence of a reliable and easily accessible ROS detection methodology applicable to platelets

A novel flow cytometry assay using dihydroethidium as redox-sensitive probe reveals NADPH oxidase-dependent generation of superoxide anion in human platelets exposed to amyloid peptide β

Reactive oxygen species (ROS) generation is critical in the regulation of platelets, which has important implications in the modulation of hemostasis and thrombosis. Nonetheless, despite several assays have been described and successfully utilized in the past, the analysis of ROS generation in human platelets remains challenging. Here we show that dihydroethidium (DHE) allows the characterization of redox responses upon platelet activation by physiological and pathological stimuli. In particular, the flow cytometry assay that we describe here allowed us to confirm that thrombin, collagen-related peptide (CRP) and arachidonic acid but not adenosine diphosphate (ADP) stimulate superoxide anion formation in a concentration-dependent manner. 0.1unit/ml thrombin, 3 μg/ml CRP and 30 μM arachidonic acid are commonly used to stimulate platelets in vitro and here were shown to stimulate a significant increase in superoxide anion formation. The ROS scavenger N-acetylcysteine (NAC) abolished superoxide anion generation in response to all tested stimuli, but the pan-NADPH oxidase (NOX) inhibitor VAS2870 only inhibited superoxide anion formation in response to thrombin and CRP. The involvement of NOXs in thrombin and CRP-dependent responses was confirmed by the inhibition of platelet aggregation induced by these stimuli by VAS2870, while platelet aggregation in response to arachidonic acid was insensitive to this inhibitor. In addition, the pathological platelet stimulus amyloid β (Aβ) 1–42 peptide induced superoxide anion formation in a concentration-dependent manner. Aβ peptide stimulated superoxide anion formation in a NOX-dependent manner, as proved by the use of VAS2870. Aβ 1–42 peptide displayed only moderate activity as an aggregation stimulus, but was able to significantly potentiate platelet aggregation in response to submaximal agonists concentrations, such as 0.03 unit/ml thrombin and 10 μM arachidonic acid. The inhibition of NOXs by 10 μM VAS2870 abolished Aβ-dependent potentiation of platelet aggregation in response to 10 μM arachidonic acid, suggesting that the pro-thrombotic activity of Aβ peptides depends on NOX activity. Similar experiments could not be performed with thrombin or collagen, as NOXs are required for the signaling induced by these stimuli. These findings shed some new light on the pro-thrombotic activity of Aβ peptides. In summary, here we describe a novel and reliable assay for the detection of superoxide anion in human platelets. This is particularly important for the investigation of the pathophysiological role of redox stress in platelets, a field of research of increasing importance, but hindered by the absence of a reliable and easily accessible ROS detection methodology applicable to platelets

A rare association of Kimura’s disease with chronic pulmonary aspergillosis

Kimura’s disease is a rare chronic benign inflammatory condition characterised by nodules of skin and soft tissue and lymphnodes. Eosinophilic infiltration is a prominent finding. It has been reported in association with various conditions like nephrotic syndrome, bronchial asthma, ulcerative colitis and aortitis syndrome. Hereby we present a case in medical literature to the best of our knowledge in Kimura’s disease in association with chronic cavitary pulmonary aspergillosis, which has never been reported. He was treated with antifungals, after which he responded clinically, eosinophilia has subsided and he is under regular follow up. Now This case report suggest possible role of chronic aspergillosis as a cause for Kimura’s disease

Common Fixed Point Theorems under Rational Inequality

In this paper we establish common fixed point theorems for two pairs of self maps in a complete metric space by using occasionally weakly biased maps satisfying the property (E.A.) using contraction  condition involving rational expressions.  These results partially generalize  Pachpatte [10], Jeong and Rhoades [5] and Kameswari [9]. Keywords: Weakly compatible, occasionally weakly compatible, property (E.A), coincidence point, point of coincidence, common fixed point. AMS (2010) Mathematics Subject Classifications: 47H10

Cover Crop Impacts on Soil Water Status

Water is a primary concern for producers in the Great Plains; as such, research is warranted to quantify how much cover crops affect the amount of soil water available to subsequent cash crops. Cover crop mixes have been marketed as a means to conserve water in no-till cropping systems following winter wheat (Triticum aestivum L.) harvest. The objectives of this study are to quantify changes in soil profile water content in the presence of different cover crops and mixtures of increasing species complexity, to quantify their biomass productivity and quality, and to quantify the impact of cover crops on subsequent corn (Zea mays L.) yields. We hypothesized the change in soil water brought on by the cover crop treatments would be correlated to the quantity of biomass produced and the species composition, rather than mixture complexity. Soil moisture was measured using a neutron probe to a depth of 9 ft. Results from 2013–14 showed no difference in water use between cover crop mixtures and single species. Cover crops depleted the soil profile by a maximum of 3.5 in. during growth, but fallow was able to gain 0.75 in. of water during the same period. At the time of corn planting, soil moisture under all cover crops had replenished to levels at cover crop emergence, except for the brassicas, which had extracted water from deeper in the profile. Corn yields were reduced following the grass cover crops and the six-species mix. Corn yields were more closely related to the carbon:nitrogen (C:N) ratio of the cover crop residue than to profile soil moisture at corn emergence. The fact that yields were similar for corn after fallow and for corn after brassica cover crops implied that water was not the cause of yield reductions after the other cover crops